Journal: EMBO Reports

Article Title: RAGE is a key regulator of ductular reaction-mediated fibrosis during cholestasis

doi: 10.1038/s44319-024-00356-7

Figure Lengend Snippet: Reagents and tools table

Article Snippet: For multiplex IF staining, HES1 signal was amplified by TSA Biotin System and detected by Cy3 conjugated streptavidin (Vector Laboratories) diluted 1:100 in TNB blocking buffer.

Techniques: Recombinant, Staining, Plasmid Preparation, Antibody Labeling, Sequencing, Negative Control, RNAscope, Modification, Saline, Software, Transfection, Enzyme-linked Immunosorbent Assay, Microscopy, Mass Spectrometry, Real-time Polymerase Chain Reaction

Journal: Cell Metabolism

Article Title: SARS-CoV-2 infects human pancreatic β cells and elicits β cell impairment

doi: 10.1016/j.cmet.2021.05.013

Figure Lengend Snippet: SARS-CoV-2-associated receptors are expressed in pancreatic β cells (A) Representative double immunofluorescence staining of ACE2, TMPRSS2, NRP1, and TFRC with the β cell marker, insulin (INS), and α cell marker, glucagon (GLU), in the normal human pancreas, donor 1. See . (B) Quantification of ACE2, TMPRSS2, NRP1, and TFRC in β cells (INS +) and α cells (GLU +) from a normal pancreas. No statistically significant changes in ACE2 and TMPRSS2 expression were detected between β and α cells. NRP1 and TFRC expression was statistically significantly higher in β cells compared with α cells. Rabbit anti-NRP1 (Abcam, ab81321, 1:200) and mouse anti-TFRC (Thermo Fisher, # 13-6800, 1:200) were used for the experiments shown here. Error bars represent mean ± SD (~10–15 islets from the pancreas of 5 non-COVID-19 donors; see ). ∗∗ p < 0.001, one-way ANOVA with Tukey’s post-test. Each dot represents one donor. Scale bars, 5 μm (A) and 2 μm (insets). See also and and .

Article Snippet: Amplification of the ISH probes was performed the next day according to manufacturer’s protocol (323100, Bio-Techne), with the final deposition of Cyanine 3 for SARS-CoV-2 spike mRNA probe targets (NEL744001KT, Akoya Biosciences).

Techniques: Double Immunofluorescence Staining, Marker, Expressing

Journal: Cell Metabolism

Article Title: SARS-CoV-2 infects human pancreatic β cells and elicits β cell impairment

doi: 10.1016/j.cmet.2021.05.013

Figure Lengend Snippet: SARS-CoV-2 preferentially infects β cells of human pancreatic islets ex vivo (A–D) Mock-treated or SARS-CoV-2-infected human pancreatic islets were stained after 2 or 6 dpi. (A) Representative double immunofluorescence staining of SARS-CoV-2 nucleocapsid protein (NP) in combination with β cell marker, insulin (INS); ɑ cell marker, glucagon (GLU); δ cell marker, somatostatin (SST); and endothelial cell marker (CD31). (B) Representative double immunofluorescence staining of SARS-CoV-2 spike protein (SP) in combination with a similar combination of markers as (A). The nuclei were stained using DAPI (blue) as a counterstain. (C) Quantified percentages of SARS-CoV-2 NP and SP within α, β, δ, and endothelial cells of pancreatic islets. Around 40% to 60% NP and SP staining, respectively, are present within β cells. (D) Quantified percentages of SARS-CoV-2 NP- and SP-positive α, β, δ, and endothelial cells. (C and D) Error bars represent mean ± SD (~500–1,000 cells were quantified from healthy isolated human islets from donors 1–5; see ). (E) Representative double immunofluorescence staining of SARS-CoV-2 NP in combination with insulin after pre-treating islets with dimethyl sulfoxide (DMSO) or 100 μM EG00229 for 1 h before infection with SARS-CoV-2. Islets were fixed at 2 dpi and stained for SARS-CoV-2 NP and β cell marker, insulin (INS). Quantification of the percentages of β cells containing NP-positive β cells (right). Error bars represent mean ± SD (~500–1,000 cells were quantified from healthy isolated human islets from donors 10–13; see ). ∗ p < 0.05, two-tailed Student’s t test. Each dot represents one donor. Scale bars, 5 μm (A, B, and E) and 2 μm (insets). See also .

Article Snippet: Amplification of the ISH probes was performed the next day according to manufacturer’s protocol (323100, Bio-Techne), with the final deposition of Cyanine 3 for SARS-CoV-2 spike mRNA probe targets (NEL744001KT, Akoya Biosciences).

Techniques: Ex Vivo, Infection, Staining, Double Immunofluorescence Staining, Marker, Isolation, Two Tailed Test

Journal: Cell Metabolism

Article Title: SARS-CoV-2 infects human pancreatic β cells and elicits β cell impairment

doi: 10.1016/j.cmet.2021.05.013

Figure Lengend Snippet: SARS-CoV-2 infects pancreatic β cells of patients with COVID-19 (A) Representative double immunofluorescence staining of pancreatic islets from patients with COVID-19 and healthy controls using antibodies against SARS-CoV-2 NP and INS. (B) Representative multiplexed images of in situ hybridization against the SARS-CoV-2 spike mRNA, in combination with immunofluorescence staining of insulin (INS). SARS-CoV-2 spike mRNA expression (red dots) was detected within pancreatic β cells. The nuclei were stained using DAPI (blue) as a counterstain. Scale bars, 5 μm (A and B) and 2 μm (insets). See also Figure S3 and .

Article Snippet: Amplification of the ISH probes was performed the next day according to manufacturer’s protocol (323100, Bio-Techne), with the final deposition of Cyanine 3 for SARS-CoV-2 spike mRNA probe targets (NEL744001KT, Akoya Biosciences).

Techniques: Double Immunofluorescence Staining, In Situ Hybridization, Immunofluorescence, Staining, Expressing

Journal: Cell Metabolism

Article Title: SARS-CoV-2 infects human pancreatic β cells and elicits β cell impairment

doi: 10.1016/j.cmet.2021.05.013

Figure Lengend Snippet: SARS-CoV-2 infection interferes with insulin content/secretion and induces β cell apoptosis (A–F) Pancreatic islet functionality was analyzed by insulin content, glucose-stimulated insulin secretion (GSIS), and TUNEL staining ex vivo . (A) Insulin content is decreased in SARS-CoV-2-infected islets compared with mock-treated islets. (B) GSIS is decreased in SARS-CoV-2-infected islets compared with mock-treated islets. (A and B) Error bars represent mean ± SD (data were collected from 7 healthy isolated human islets, donors 2–8; see ). ∗ p < 0.05, two-tailed Student’s t test. (C) Representative staining of β cell apoptosis by in situ TUNEL and DAPI staining in β cells (INS) of mock- or SARS-CoV-2-treated human islets. DNase-treated sections were used as a positive control in the TUNEL assay. (D and F) Quantification of the percentages of islets containing TUNEL-positive β cells. Error bars represent mean ± SD (~500–1,000 cells were quantified from each of 3–5 separate healthy isolated human islets, donors 1–5 [D] and 7–9 [F]; see ). (E) Representative staining of β cell apoptosis by in situ TUNEL and DAPI staining in β cells (INS) of mock-treated versus SARS-CoV-2-SP-treated human islets. ∗ p < 0.05, ∗∗ p < 0.01, two-tailed Student’s t test. Scale bars, 5 μm (C and E). See also and , , , and .

Article Snippet: Amplification of the ISH probes was performed the next day according to manufacturer’s protocol (323100, Bio-Techne), with the final deposition of Cyanine 3 for SARS-CoV-2 spike mRNA probe targets (NEL744001KT, Akoya Biosciences).

Techniques: Infection, TUNEL Assay, Staining, Ex Vivo, Isolation, Two Tailed Test, In Situ, Positive Control

Journal: Cell Metabolism

Article Title: SARS-CoV-2 infects human pancreatic β cells and elicits β cell impairment

doi: 10.1016/j.cmet.2021.05.013

Figure Lengend Snippet:

Article Snippet: Amplification of the ISH probes was performed the next day according to manufacturer’s protocol (323100, Bio-Techne), with the final deposition of Cyanine 3 for SARS-CoV-2 spike mRNA probe targets (NEL744001KT, Akoya Biosciences).

Techniques: Virus, Recombinant, Blocking Assay, Enzyme-linked Immunosorbent Assay, In Situ, Control, RNAscope, Multiplex Assay, Purification, Expressing, Infection, Software